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  本文選題：DNA甲基化 + 基因表達��； 參考：《暨南大學》2009年碩士論文

【摘要】： 目的:通過比較不同細胞類型胰島組織特異性基因DNA甲基化程度差異及與基因表達的關系,探討DNA甲基化在胰島組織特異性基因表達和胰島分化中的作用。 方法:采用甲基化DNA免疫共沉淀—實時定量PCR法(MeDIP—qPCR)比較小鼠胰島素瘤β細胞(NIT1)、NIH小鼠成纖維細胞(NIH3T3)及小鼠胚胎干細胞(mES)三者Pdx-1、Pax4、MafA、Nkx6.1和Insulin等基因轉錄起始區(qū)DNA甲基化程度差異。同時采用實時定量RT-PCR檢測上述三種細胞各基因mRNA表達水平。比較這些基因DNA甲基化水平與基因表達之間的相互關系。 結果:NIH3T3細胞Pdx-1、MafA和Nkx6.1基因轉錄起始區(qū)呈高甲基化,在NIT1細胞和mES細胞中這些基因則呈低甲基化,前者的甲基化程度明顯高于后兩者(P＜0.05)。另外兩種基因Pax4和Insulin轉錄起始區(qū)在三種細胞中呈低甲基化,NIH3T3細胞Pax4基因的甲基化程度與NIT1細胞無統(tǒng)計學差異(P＞0.05);前者Insulin基因呈非甲基化,其甲基化程度低于后者(P＜0.05)。在基因表達方面,NIT1細胞高表達Pdx-1、Pax4、MafA、Nkx6.1和Insulin等基因,NIH3T3和mES細胞則未見這些基因表達。結果顯示高甲基化狀態(tài)的NIH3T3細胞無Pdx-1、MafA和Nkx6.1表達,而呈低甲基化狀態(tài)的NIT1細胞則高表達Pdx-1、MafA和Nkx6.1。Pax4和Insulin基因甲基化狀態(tài)與基因表達之間則無相關性。在胚胎干細胞中各基因呈低甲基化狀態(tài),也未見基因表達。 結論:DNA甲基化參與了胰島組織特異性基因的表達調控,且在胚胎干細胞向胰島分化的過程中起重要調節(jié)作用。
[Abstract]:Aim: to investigate the role of DNA methylation in islet tissue specific gene expression and islet differentiation by comparing the degree of DNA methylation of islet specific genes in different cell types and the relationship between DNA methylation and gene expression.At the same time, real-time quantitative RT-PCR was used to detect the mRNA expression level of the three kinds of cells.To compare the correlation between DNA methylation level and gene expression of these genes.Results the transcription initiation region of Pdx-1mafA and Nkx6.1 gene was hypermethylated in the cell line of: NIH3T3, but it was hypomethylated in NIT1 cells and mES cells. The methylation degree of these genes in the former was significantly higher than that in the latter two cells (P < 0.05).There was no significant difference in the degree of methylation of the Pax4 gene between the two genes (Pax4 and Insulin) in the three cells (P > 0.05), but the former was demethylated and the degree of methylation was lower than that of the latter (P < 0.05).The expression of these genes in NIT1 cells was not found in NIH3T3 and mES cells.The results showed that there was no expression of Pdx-1mafA and Nkx6.1 in hypermethylated NIH3T3 cells, but there was no correlation between the methylation status of Pdx-1MafA and Nkx6.1.Pax4 and Insulin gene expression in NIT1 cells with low methylation state.The genes in embryonic stem cells were hypomethylated and no gene expression was found.Conclusion DNA methylation is involved in the regulation of islet tissue specific gene expression and plays an important role in the differentiation of embryonic stem cells into pancreatic islets.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R657.5;R346

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本文編號：1735569